RNA-seq profiles of Madurella mycetomatis mm55 cultured pathogen

Mycetoma is a neglected, chronic granulomatous infection of the subcutaneous tissue, most often caused by the fungal pathogen Madurella mycetomatis. Characteristic of the infection is the formation of grains. M. mycetomatis genome strain MM55 was cultured on Sabouraud Dextrose Agar for two to three weeks at 37⁰C. Mycelium was harvested, sonicated at 28 micron (Soniprep 150, Beun de Ronde, the Netherlands) and incubated at 37⁰C in RPMI-media supplemented with 0.35 g/L L-glutamine, 1.98 mM 4-morpholinepropanesulfonic acid (MOPS) and 100 mg/L chloramphenicol. After two weeks, the mycelia were separated and washed by vacuum filtration (Nalgene, Abcoude, the Netherlands using a 22 micron filter (Whatman). The mycelial biomass was scraped from the filter, the wet weight was determined and a suspension was prepared containing 100 mg mycelial biomass per ml phosphate-buffered saline (PBS). The suspension was sonicated at 28 micron for 2 minutes (Soniprep 150, Beun de Ronde, the Netherlands), the resulting homogeneous suspension was washed once in PBS and again diluted to a concentration of 4 mg per 40 µL PBS, corresponding to 600-850 CFU/larvae.